Isolation and mass spectrometry of transcription factor complexes Journal Article

Authors: Sebastiaan Winkler, G.; Lacomis, L.; Philip, J.; Erdjument-Bromage, H.; Svejstrup, J. Q.; Tempst, P.
Article Title: Isolation and mass spectrometry of transcription factor complexes
Abstract: Protocols are described that enable the isolation of novel proteins associated with a known protein and the subsequent identification of these proteins by mass spectrometry. We review the basics of nanosample handling and of two complementary approaches to mass analysis, and provide protocols for the entire process. The protein isolation procedure is rapid and based on two high-affinity chromatography steps. The method does not require previous knowledge of complex composition or activity and permits subsequent biochemical characterization of the isolated factor. As an example, we provide the procedures used to isolate and analyze yeast Elongator, a histone acetyltransferase complex important for transcript elongation, which led to the identification of three novel subunits. © 2002 Elsevier Science (USA). All rights reserved.
Keywords: controlled study; protein expression; unclassified drug; nonhuman; molecular genetics; methodology; mass spectrometry; protein analysis; metabolism; protein binding; transcription factor; transcription factors; chemistry; amino acid sequence; molecular sequence data; protein purification; saccharomyces cerevisiae; nucleotide sequence; peptide fragments; peptide fragment; base sequence; saccharomyces cerevisiae proteins; protein subunit; saccharomyces cerevisiae protein; protein subunits; isolation and purification; fungal protein; acyltransferase; acetyltransferases; macromolecule; macromolecular substances; protein isolation; affinity chromatography; chromatography, affinity; histone acetyltransferase; histone acetyltransferases; priority journal; article; support, non-u.s. gov't; protein elp1; macromolecular systems; microchemistry; spectrum analysis, mass
Journal Title: Methods
Volume: 26
Issue: 3
ISSN: 1046-2023
Publisher: Academic Press Inc., Elsevier Science  
Date Published: 2002-03-01
Start Page: 260
End Page: 269
Language: English
DOI: 10.1016/s1046-2023(02)00030-0
PUBMED: 12054882
PROVIDER: scopus
Notes: Export Date: 14 November 2014 -- Source: Scopus
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MSK Authors
  1. Paul J Tempst
    307 Tempst
  2. John Philip
    20 Philip
  3. Lynne M Lacomis
    21 Lacomis